The vertical distribution of magnetotactic bacteria along various physico-chemical gradients in freshwater microcosms was analyzed by a combined approach of viable cell counts, 16S rRNA gene analysis, microsensor profiling and biogeochemical methods. The occurrence of magnetotactic bacteria was restricted to a narrow sediment layer overlapping or closely below the maximum oxygen and nitrate penetration depth. Different species showed different preferences within vertical gradients, but the largest proportion (63-98%) of magnetotactic bacteria was detected within the suboxic zone. In one microcosm the community of magnetotactic bacteria was dominated by one species of a coccoid "Alphaproteobacterium", as detected by denaturing gradient gel electrophoresis in sediment horizons from 1 to 10 mm depth. Maximum numbers of magnetotactic bacteria were up to 1.5 x 10(7) cells/cm3, which corresponded to 1% of the total cell number in the upper sediment layer. The occurrence of magnetotactic bacteria coincided with the availability of significant amounts (6-60 microM) of soluble Fe(II), and in one sample with hydrogen sulfide (up to 40 microM). Although various trends were clearly observed, a strict correlation between the distribution of magnetotactic bacteria and individual geochemical parameters was absent. This is discussed in terms of metabolic adaptation of various strains of magnetotactic bacteria to stratified sediments and diversity of the magnetotactic bacterial communities.

Both magnetic collection and "race track" purification techniques were highly effective for selective enrichment of magnetotactic bacteria (MTB) from complex communities, as suggested by amplified ribosomal DNA restriction analysis and denaturing gradient gel electrophoresis combined with sequence analysis of 16S rRNA genes. Using these purification methods, the occurrence and diversity of MTB in microcosms from various marine and freshwater environments were assayed by using a combined microscopic, molecular, and cultivation approach. Most microcosms were dominated by magnetotactic cocci. Consistently, the majority of retrieved 16S RNA sequences were affiliated with a distinct cluster in the Alphaproteobacteria. Within this lineage the levels of sequence divergence were <1 to 11%, indicating genus-level diversity between magnetotactic cocci from various microcosms, as well as between MTB from different stages of succession of the same microcosms. The community composition in microscosms underwent drastic succession during incubation, and significant heterogeneities were observed between microcosms from the same environmental sources. A novel magnetotactic rod (MHB-1) was detected in a sediment sample from a lake in northern Germany by fluorescence in situ hybridization. MHB-1 falls into the Nitrospira phylum, displaying 91% 16S rRNA sequence similarity to "Magnetobacterium bavaricum." In extensive cultivation attempts, we failed to isolate MHB-1, as well as most other MTB present in our samples. However, although magnetotactic spirilla were not frequently observed in the enrichments, 10 novel isolates of the genus Magnetospirillum which had not routinely been isolated in pure culture before were obtained.Magnetotactic bacteria (MTB) are a heterogeneous group of aquatic microorganisms which share the ability to orient themselves along magnetic field lines. Magnetic orientation is due to the presence of magnetosomes, which are intracellular membrane-bound crystals of magnetic iron mineral which consist of magnetite or greigite (3,28,29). Diverse MTB, including cocci, spirilla, rods, vibrios, and multicellular aggregates, have been found in different aquatic habitats (5,7,17). Because of their high abundance and their remarkable potential to accumulate and precipitate iron minerals, MTB are assumed to have great impact in the biogeochemical cycling in natural sediments, which, however, has remained poorly understood. Most cultivated and uncultivated MTB have been affiliated with the Alphaproteobacteria. Moreover, the magnetotactic sulfate-reducing bacterium Desulfovibrio magneticus (14) and a magnetotactic, many-celled prokaryote (9) belong to the Deltaproteobacteria, while a giant magnetotactic rod, tentatively named "Magnetobacterium bavaricum," is affiliated with the Nitrospira phylum (34). Recently, a greigite-producing rod belonging to the ␥-Proteobacteria was identified (32). The phylogenetic and morphological diversity of MTB is matched by notable variation in the shapes, organizations, and numbers of magne...

In this report, we describe the selective cloning of large DNA fragments from magnetotactic metagenomes from various aquatic habitats. This was achieved by a two-step magnetic enrichment which allowed the mass collection of environmental magnetotactic bacteria (MTB) virtually free of nonmagnetic contaminants. Four fosmid libraries were constructed and screened by end sequencing and hybridization analysis using heterologous magnetosome gene probes. A total of 14 fosmids were fully sequenced. We identified and characterized two fosmids, most likely originating from two different alphaproteobacterial strains of MTB that contain several putative operons with homology to the magnetosome island (MAI) of cultivated MTB. This is the first evidence that uncultivated MTB exhibit similar yet differing organizations of the MAI, which may account for the diversity in biomineralization and magnetotaxis observed in MTB from various environments.

Desulfovibrio magneticus strain RS-l is an anaerobic sulfate-reducing bacterium. Cells form intracellular nanocrystals of magnetite but are only weakly magnetotactic. In order to understand the unusual magnetic response of this strain, we studied magnetite crystals within cells grown with fumarate and sulfate. Many cells grown under either condition did not form magnetic crystals while others contained only 1 to 18 small (~40 nm) magnetite-containing magnetosomes. Bulk magnetic measurements of whole cells showed a superparamagnetic-like behavior, indicating that many of the magnetite crystals are too small to have a permanent magnetic moment at ambient temperature. The temperature of the Verwey transition is lower (~ 86 K) than of magnetite from other magnetotactic strains, likely indicating partial oxidation of magnetite into maghemite. As a result of the small size and small number of magnetite magnetosomes, the magnetic moments of most cells grown anaerobically with fumarate or sulfate are insufficient for magnetotaxis.In addition to intracellular magnetite, in some cultures another iron oxide, hematite, formed on the surfaces of cells. The hematite grains are embedded in an extracellular polymeric material, indicating that the crystals likely resulted from a biologically induced mineralization process. Although the hematite particles appear to consist of aggregations of many small (5 to 10 nm) grains, the grains have a consensus orientation and thus the whole particle diffracts as a single crystal. The aligned arrangement of nanoparticles within larger clusters may reflect either a templated nucleation of hematite crystallites in an extracellular organic matrix, or result from a self-assembling process during the crystallization of hematite from ferric gels or ferrihydrite.

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